DNA encoding parathyroid hormone receptor

Abstract

DNA encoding a parathyroid hormone receptor; production and isolation of recombinant and synthetic parathyroid hormone receptor polypeptides and fragments; antibodies to parathyroid hormone receptors and receptor fragments; methods for screening candidate compounds for antagonistic or agonistic effects on parathyroid hormone receptor action; and diagnostic and therapeutic methods of these compounds are disclosed.

1. An isolated DNA comprising a nucleic acid sequence that encodes a polypeptide comprising an amino acid sequence at least 75% identical to SEQ ID NO:20, wherein the polypeptide binds parathyroid hormone or parathyroid hormone-related protein. 
     
     
       2. The isolated DNA of  claim 1 , wherein the nucleic acid sequence encodes substantially all of SEQ ID NO: 18. 
     
     
       3. The isolated DNA of  claim 1 , wherein the nucleic acid sequence encodes substantially all of SEQ ID NO: 19. 
     
     
       4. The isolated DNA of  claim 1 , wherein the nucleic acid sequence encodes substantially all of SEQ ID NO: 20. 
     
     
       5. The isolated DNA of  claim 1 , wherein the nucleic acid sequence encodes substantially all of SEQ ID NO: 21. 
     
     
       6. A cell in culture containing the isolated DNA of  claim 1 . 
     
     
       7. An essentially homogenous population of cells in culture, each of which comprises the isolated DNA of  claim 1 . 
     
     
       8. The isolated DNA of  claim 1 , wherein the DNA is single-stranded. 
     
     
       9. The single-stranded isolated DNA of  claim 8 , wherein the DNA is detectably labeled. 
     
     
       10. A vector comprising the isolated DNA of  claim 1 . 
     
     
       11. A method of producing a polypeptide, the method comprising the steps of:
 (a) providing a cell comprising the isolated DNA of  claim 1 ; and 
 (b) culturing the cell under conditions that permit expression of a polypeptide from the isolated DNA, to thereby produce the polypeptide. 
 
     
     
       12. A cell in culture containing the isolated DNA of  claim 5 . 
     
     
       13. An essentially homogenous population of cells in culture, each of which comprises the isolated DNA of  claim 5 . 
     
     
       14. The isolated DNA of  claim 5 , wherein the DNA is single-stranded. 
     
     
       15. The single-stranded isolated DNA of  claim 14 , wherein the DNA is detectably labeled. 
     
     
       16. A vector comprising the isolated DNA of  claim 5 . 
     
     
       17. A method of producing a polypeptide, the method comprising the steps of:
 (a) providing a cell comprising the isolated DNA of  claim 5 ; and 
 (b) culturing the cell under conditions that permit expression of a polypeptide from the isolated DNA, to thereby produce the polypeptide. 
 
     
     
       18. An isolated DNA comprising a sequence that hybridizes under conditions of high stringency to a probe the sequence of which consists of the coding sequence of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, or a sequence complementary to the coding sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, wherein the DNA encodes a polypeptide that binds to parathyroid hormone or parathyroid hormone-related protein, and wherein conditions of high stringency include hybridizing at 42° C. for 18–24 h in a solution comprising 50% formamide, 4× saline sodium citrate (SSC), 2× Denhardt's solution, 10% Dextran sulphate and 100 μg/ml salmon sperm DNA and washing with 2×SSC/0.1% SDS for 30 minutes at room temperature and then with 1×SSC/0.1% SDS for 30 minutes at 45° C. 
     
     
       19. A cell in culture containing the isolated DNA of  claim 18 . 
     
     
       20. An essentially homogenous population of cells in culture, each of which comprises the isolated DNA of  claim 18 . 
     
     
       21. The isolated DNA of  claim 18 , wherein the DNA is single-stranded. 
     
     
       22. The single-stranded isolated DNA of  claim 21 , wherein the DNA is detectably labeled. 
     
     
       23. A vector comprising the isolated DNA of  claim 18 . 
     
     
       24. A method of producing a polypeptide, the method comprising the steps of:
 (a) providing a cell comprising the isolated DNA of  claim 18 ; and 
 (b) culturing the cell under conditions that permit expression of a polypeptide from the isolated DNA, to thereby produce the polypeptide. 
 
     
     
       25. An isolated DNA comprising a sequence that hybridizes under conditions of high stringency to a probe the sequence of which consists of the coding sequence of SEQ ID NO: 4, or a sequence complementary to the coding sequence of SEQ ID NO: 4, wherein the DNA encodes a polypeptide that binds to parathyroid hormone or parathyroid hormone-related protein, and wherein conditions of high stringency include hybridizing at 42° C. for 18–24 h in a solution comprising 50% formamide, 4× saline sodium citrate (SSC), 2× Denhardt's solution, 10% Dextran sulphate and 100 μg/ml salmon sperm DNA and washing with 2×SSC/0.1% SDS for 30 minutes at room temperature and then with 1×SSC/0.1% SDS for 30 minutes at 45° C. 
     
     
       26. A cell in culture containing the isolated DNA of  claim 25 . 
     
     
       27. An essentially homogenous population of cells in culture, each of which comprises the isolated DNA of  claim 25 . 
     
     
       28. The isolated DNA of  claim 25 , wherein the DNA is single-stranded. 
     
     
       29. The single-stranded isolated DNA of  claim 28 , wherein the DNA is detectably labeled. 
     
     
       30. A vector comprising the isolated DNA of  claim 25 . 
     
     
       31. A method of producing a polypeptide, the method comprising the steps of:
 (a) providing a cell comprising the isolated DNA of  claim 25 ; and 
 (b) culturing the cell under conditions that permit expression of a polypeptide from the isolated DNA, to thereby produce the polypeptide. 
 
     
     
       32. An isolated DNA comprising a nucleotide sequence encoding a polypeptide comprising SEQ ID NO:18, SEQ ID NO:19 or SEQ ID NO:20. 
     
     
       33. The isolated DNA of  claim 32 , wherein the nucleotide sequence comprises the coding sequence of SEQ ID NO:1. 
     
     
       34. The isolated DNA of  claim 32 , wherein the nucleotide sequence comprises the coding sequence of SEQ ID NO:2. 
     
     
       35. The isolated DNA of  claim 32 , wherein the nucleotide sequence comprises the coding sequence of SEQ ID NO: 3. 
     
     
       36. A cell in culture containing the isolated DNA of  claim 32 . 
     
     
       37. An essentially homogenous population of cells in culture, each of which comprises the isolated DNA of  claim 32 . 
     
     
       38. The isolated DNA of  claim 32 , wherein the DNA is single-stranded. 
     
     
       39. The single-stranded isolated DNA of  claim 38 , wherein the DNA is detectably labeled. 
     
     
       40. A vector comprising the isolated DNA of  claim 32 . 
     
     
       41. A method of producing a polypeptide, the method comprising the steps of:
 (a) providing a cell comprising the isolated DNA of  claim 32 ; and 
 (b) culturing the cell under conditions that permit expression of a polypeptide from the isolated DNA, to thereby produce the polypeptide. 
 
     
     
       42. An isolated DNA comprising a nucleotide sequence encoding a polypeptide comprising SEQ ID NO: 21. 
     
     
       43. The isolated DNA of  claim 42 , wherein the nucleotide sequence comprises the coding sequence of SEQ ID NO:4. 
     
     
       44. A cell in culture containing the isolated DNA of  claim 42 . 
     
     
       45. An essentially homogenous population of cells in culture, each of which comprises the isolated DNA of  claim 42 . 
     
     
       46. The isolated DNA of  claim 42 , wherein the DNA is single-stranded. 
     
     
       47. The single-stranded isolated DNA of  claim 46 , wherein the DNA is detectably labeled. 
     
     
       48. A vector comprising the isolated DNA of  claim 42 . 
     
     
       49. A method of producing a polypeptide, the method comprising the steps of:
 (a) providing a cell comprising the isolated DNA of  claim 42 ; and 
 (b) culturing the cell under conditions that permit expression of a polypeptide from the isolated DNA, to thereby produce the polypeptide. 
 
     
     
       50. An isolated DNA comprising a nucleotide sequence encoding a polypeptide consisting of SEQ ID NO:18, SEQ ID NO:19 or SEQ ID NO:20. 
     
     
       51. A cell in culture containing the isolated DNA of  claim 42 . 
     
     
       52. An essentially homogenous population of cells in culture, each of which comprises the isolated DNA of  claim 50 . 
     
     
       53. The isolated DNA of  claim 50 , wherein the DNA is single-stranded. 
     
     
       54. The single-stranded isolated DNA of  claim 53 , wherein the DNA is detectably labeled. 
     
     
       55. A vector comprising the isolated DNA of  claim 50 . 
     
     
       56. A method of producing a polypeptide, the method comprising the steps of:
 (a) providing a cell comprising the isolated DNA of  claim 50 ; and 
 (b) culturing the cell under conditions that permit expression of a polypeptide from the isolated DNA, to thereby produce the polypeptide. 
 
     
     
       57. An isolated DNA comprising a nucleotide sequence encoding a polypeptide consisting of SEQ ID NO:21. 
     
     
       58. A cell in culture containing the isolated DNA of  claim 57 . 
     
     
       59. An essentially homogenous population of cells in culture, each of which comprises the isolated DNA of  claim 57 . 
     
     
       60. The isolated DNA of  claim 57 , wherein the DNA is single-stranded. 
     
     
       61. The single-stranded isolated DNA of  claim 60 , wherein the DNA is detectably labeled. 
     
     
       62. A vector comprising the isolated DNA of  claim 57 . 
     
     
       63. A method of producing a polypeptide, the method comprising the steps of:
 (a) providing a cell comprising the isolated DNA of  claim 57 ; and 
 (b) culturing the cell under conditions that permit expression of a polypeptide from the isolated DNA, to thereby produce the polypeptide. 
 
     
     
       64. An isolated DNA comprising a nucleotide sequence encoding a polypeptide comprising an extracellular domain of SEQ ID NO:18, SEQ ID NO:19 or SEQ ID NO:20. 
     
     
       65. A cell in culture containing the isolated DNA of  claim 64 . 
     
     
       66. An essentially homogenous population of cells in culture, each of which comprises the isolated DNA of  claim 64 . 
     
     
       67. The isolated DNA of  claim 64 , wherein the DNA is single-stranded. 
     
     
       68. The single-stranded isolated DNA of  claim 67 , wherein the DNA is detectably labeled. 
     
     
       69. A vector comprising the isolated DNA of  claim 64 . 
     
     
       70. A method of producing a polypeptide, the method comprising the steps of:
 (a) providing a cell comprising the isolated DNA of  claim 64 ; and 
 (b) culturing the cell under conditions that permit expression of a polypeptide from the isolated DNA, to thereby produce the polypeptide. 
 
     
     
       71. An isolated DNA comprising a nucleotide sequence encoding a polypeptide comprising an extracellular domain of SEQ ID NO:21. 
     
     
       72. A cell in culture containing the isolated DNA of  claim 71 . 
     
     
       73. An essentially homogenous population of cells in culture, each of which comprises the isolated DNA of  claim 71 . 
     
     
       74. The isolated DNA of  claim 71 , wherein the DNA is single-stranded. 
     
     
       75. The single-stranded isolated DNA of  claim 74 , wherein the DNA is detectably labeled. 
     
     
       76. A vector comprising the isolated DNA of  claim 71 . 
     
     
       77. A method of producing a polypeptide, the method comprising the steps of:
 (a) providing a cell comprising the isolated DNA of  claim 71 ; and 
 (b) culturing the cell under conditions that permit expression of a polypeptide from the isolated DNA, to thereby produce the polypeptide. 
 
     
     
       78. An isolated DNA comprising a nucleotide sequence encoding a polypeptide comprising SEQ ID NO:18, SEQ ID NO:19 or SEQ ID NO:20 with at least one conservative amino acid substitution, wherein the polypeptide binds parathyroid hormone or parathyroid hormone-related protein. 
     
     
       79. A cell in culture containing the isolated DNA of  claim 78 . 
     
     
       80. An essentially homogenous population of cells in culture, each of which comprises the isolated DNA of  claim 78 . 
     
     
       81. The isolated DNA of  claim 78 , wherein the DNA is single-stranded. 
     
     
       82. The single-stranded isolated DNA of  claim 81 , wherein the DNA is detectably labeled. 
     
     
       83. A vector comprising the isolated DNA of  claim 78 . 
     
     
       84. A method of producing a polypeptide, the method comprising the steps of:
 (a) providing a cell comprising the isolated DNA of  claim 78 ; and 
 (b) culturing the cell under conditions that permit expression of a polypeptide from the isolated DNA, to thereby produce the polypeptide. 
 
     
     
       85. An isolated DNA comprising a nucleotide sequence encoding a polypeptide comprising SEQ ID NO:21 with at least one conservative amino acid substitution, wherein the polypeptide binds parathyroid hormone or parathyroid hormone-related protein. 
     
     
       86. A cell in culture containing the isolated DNA of  claim 85 . 
     
     
       87. An essentially homogenous population of cells in culture, each of which comprises the isolated DNA of  claim 85 . 
     
     
       88. The isolated DNA of  claim 85 , wherein the DNA is single-stranded. 
     
     
       89. The single-stranded isolated DNA of  claim 88 , wherein the DNA is detectably labeled. 
     
     
       90. A vector comprising the isolated DNA of  claim 85 . 
     
     
       91. A method of producing a polypeptide, the method comprising the steps of:
 (a) providing a cell comprising the isolated DNA of  claim 85 ; and 
 (b) culturing the cell under conditions that permit expression of a polypeptide from the isolated DNA, to thereby produce the polypeptide.